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R&D Systems human recombinant scd14

Biodistribution of intact (>30 kDa, black) and degraded (<30 kDa, white) [ 14 <t>C]sCD14</t> and a digestible control protein, [ 14 C]BSA, postingestion in 10 d old rats. A, stomach; B, duodenum; C, jejunum; D, ileum; E, large intestine. Rat pups were gavage fed 25 μg of [ 14 <t>C]sCD14</t> (1.63 ×10 5 dpm/μg) or [ 14 C]BSA (3.44 ×10 4 dpm/μg). Post-euthanasia (0.3, 4 or 8 h), proteins were extracted from organs, size separated using a 30 kDa cut-off spin column and liquid scintillation counted. Data are presented as mean ± SD, and n=3 for each time point. * P <0.05, using a Student’s t -test.

Human Recombinant Scd14, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/recombinant+human+scd14/Recombinant+Human+CD14+Protein/pmc04965269-114-0-3
Average 93 stars, based on 40 article reviews

human recombinant scd14 - by Bioz Stars, 2026-11

93/100 stars

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1) Product Images from "Ingested soluble CD14 from milk is transferred intact into the blood of newborn rats"

Article Title: Ingested soluble CD14 from milk is transferred intact into the blood of newborn rats

Journal: Pediatric research

doi: 10.1038/pr.2013.225

Biodistribution of intact (>30 kDa, black) and degraded (<30 kDa, white) [ 14 C]sCD14 and a digestible control protein, [ 14 C]BSA, postingestion in 10 d old rats. A, stomach; B, duodenum; C, jejunum; D, ileum; E, large intestine. Rat pups were gavage fed 25 μg of [ 14 C]sCD14 (1.63 ×10 5 dpm/μg) or [ 14 C]BSA (3.44 ×10 4 dpm/μg). Post-euthanasia (0.3, 4 or 8 h), proteins were extracted from organs, size separated using a 30 kDa cut-off spin column and liquid scintillation counted. Data are presented as mean ± SD, and n=3 for each time point. * P <0.05, using a Student’s t -test.

Figure Legend Snippet: Biodistribution of intact (>30 kDa, black) and degraded (<30 kDa, white) [ 14 C]sCD14 and a digestible control protein, [ 14 C]BSA, postingestion in 10 d old rats. A, stomach; B, duodenum; C, jejunum; D, ileum; E, large intestine. Rat pups were gavage fed 25 μg of [ 14 C]sCD14 (1.63 ×10 5 dpm/μg) or [ 14 C]BSA (3.44 ×10 4 dpm/μg). Post-euthanasia (0.3, 4 or 8 h), proteins were extracted from organs, size separated using a 30 kDa cut-off spin column and liquid scintillation counted. Data are presented as mean ± SD, and n=3 for each time point. * P <0.05, using a Student’s t -test.

Techniques Used: Control

The biodistribution of [ 125 I]sCD14 (A) and a digestible control protein, [ 125 I]BSA (B), post-ingestion in 10 d old rats. Rat pups were gavage fed 25 μg of [ 125 I]sCD14 (12.16 ×10 6 dpm/μg) or [ 125 I]BSA (13.7 ×10 6 dpm/μg). Post-euthanasia (0.3, 4 or 8 h), proteins were extracted from harvested organs, size separated by SDS-PAGE and phosphor-imaged. Stomach (St) and contents (St Con), duodenum (Duo) and contents (Duo Con), jejunum (Je) and contents (Je Con), ileum and contents (Ileum Con), large intestine (L In) and contents (L In Con), liver and blood. Figure is representative of n=3.

Figure Legend Snippet: The biodistribution of [ 125 I]sCD14 (A) and a digestible control protein, [ 125 I]BSA (B), post-ingestion in 10 d old rats. Rat pups were gavage fed 25 μg of [ 125 I]sCD14 (12.16 ×10 6 dpm/μg) or [ 125 I]BSA (13.7 ×10 6 dpm/μg). Post-euthanasia (0.3, 4 or 8 h), proteins were extracted from harvested organs, size separated by SDS-PAGE and phosphor-imaged. Stomach (St) and contents (St Con), duodenum (Duo) and contents (Duo Con), jejunum (Je) and contents (Je Con), ileum and contents (Ileum Con), large intestine (L In) and contents (L In Con), liver and blood. Figure is representative of n=3.

Techniques Used: Control, SDS Page

Uptake of intact (A, >30 kDa) and degraded (B, <30 kDa) [ 125 I]sCD14 into the blood of 10 d old rat pups. Data are presented as mean ± SD, and n=4. Percent dose fed >30 kDa ( A ) at 0.3 h was significantly less than all subsequent time points (*, P <0.05). Percent dose fed <30 kDa ( B ) at 0.3 h was significantly less than the percent dose fed at 4, 5 and 7 h (*, P <0.05, Student’s t -test).

Figure Legend Snippet: Uptake of intact (A, >30 kDa) and degraded (B, <30 kDa) [ 125 I]sCD14 into the blood of 10 d old rat pups. Data are presented as mean ± SD, and n=4. Percent dose fed >30 kDa ( A ) at 0.3 h was significantly less than all subsequent time points (*, P <0.05). Percent dose fed <30 kDa ( B ) at 0.3 h was significantly less than the percent dose fed at 4, 5 and 7 h (*, P <0.05, Student’s t -test).

Techniques Used:

Intactness of ingested [ 125 I]sCD14 in the blood of 10 d old rat pups post size separation (< or >30 kDa) visualized by phosphor-imaging (A) and silver staining (B). Intact and non-ingested [ 125 I]sCD14 was used as a positive control. Figure is representative of n=4.

Figure Legend Snippet: Intactness of ingested [ 125 I]sCD14 in the blood of 10 d old rat pups post size separation (< or >30 kDa) visualized by phosphor-imaging (A) and silver staining (B). Intact and non-ingested [ 125 I]sCD14 was used as a positive control. Figure is representative of n=4.

Techniques Used: Imaging, Silver Staining, Positive Control

The presence of rat-sCD14 in rat milk, 10 days postpartum, as detected by immunoblotting. Lanes: 1, molecular weight marker; 2, rat milk; 3, human recombinant sCD14 (100 ng, positive control).

Figure Legend Snippet: The presence of rat-sCD14 in rat milk, 10 days postpartum, as detected by immunoblotting. Lanes: 1, molecular weight marker; 2, rat milk; 3, human recombinant sCD14 (100 ng, positive control).

Techniques Used: Western Blot, Molecular Weight, Marker, Recombinant, Positive Control

Recombinant:

Article Title: Induction of Keratinocyte Growth Factor 1 Expression by Lipopolysaccharide Is Regulated by CD-14 and Toll-Like Receptors 2 and 4
Article Snippet: .. Cultures were treated with E. coli LPS (O55:B; Sigma catalog no. L2880) or P. gingivalis LPS (ATCC 33211) (40) at 5 to 100 ng/ml or recombinant human sCD14 (R&D Systems Inc, Minneapolis, Minn.) at 50 ng/ml with -MEM in the presence and absence of 1% FBS. .. In some experiments, cell cultures were preincubated for 30 min with specific blocking anti-human TLR2 (Cascade BioScience, Winchester, Mass.) and/or TLR4 (eBioscience, San Diego, Calif.) monoclonal antibodies at 0.5 g/ml.

Article Title: Induction of Keratinocyte Growth Factor 1 Expression by Lipopolysaccharide Is Regulated by CD-14 and Toll-Like Receptors 2 and 4
Article Snippet: .. Cultures were treated with E. coli LPS (O55:B; Sigma catalog no. L2880) or P. gingivalis LPS (ATCC 33211) ( 40 ) at 5 to 100 ng/ml or recombinant human sCD14 (R&D Systems Inc, Minneapolis, Minn.) at 50 ng/ml with α-MEM in the presence and absence of 1% FBS. .. In some experiments, cell cultures were preincubated for 30 min with specific blocking anti-human TLR2 (Cascade BioScience, Winchester, Mass.) and/or TLR4 (eBioscience, San Diego, Calif.) monoclonal antibodies at 0.5 μg/ml.

Article Title: Monitoring of endothelial cell activation in experimental sepsis with a two-step cell culture model.
Article Snippet: Cell culture media and reagents The cell culture media RPMI-1640 (RPMI), Medium 199 (M199) and Hank’s Balanced Salt Solution (HBSS), phosphate-buffered saline (PBS), ethylene diamine tetraacetic acid disodium salt (EDTA), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), penicillin-streptomycin (PS) and LPS from Pseudomonas aeruginosa were purchased from Sigma-Aldrich (St Louis, MO, USA). .. Recombinant human LPS-binding protein (LBP) and recombinant human sCD14 were from R&D Systems (Minneapolis, MN, USA). .. Phycoerythrin (PE)-conjugated mouse anti-human CD54 (anti-ICAM1) and PE-Cy5-conjugated mouse anti-human CD62E (anti-E-selectin) monoclonal antibodies were purchased from BD Biosciences (San Diego, CA, USA).

Article Title: Multi-Hit Inhibition of Circulating and Cell-Associated Components of the Toll-Like Receptor 4 Pathway by Oxidized Phospholipids
Article Snippet: .. Recombinant human sCD14 was a gift from Dr. Yoshio Kuroki, Sapporo Medical University, Japan or purchased from R&D Systems, Minneapolis, MN, USA. .. Acyclic lipid A analog E6020 (ER112066) capable of activating TLR4/MD-2 in the absence of LBP and CD14 1 was synthesized at the Eisai Research Institute (Andover, MA, USA).

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Elevated levels of proinflammatory biomarkers in LC patients. (A) Cumulative data comparing the plasma IL-1α, (B) IL-6, (C) TNF-α, (D) IP-10, (E) CRP, (F) SAA, (G) IL-13, and (H) IL-1β, (I) Flt-1, and (J) soluble CD14 measured by ELISA in the plasma of R, LC, and HC groups. (K) Comparing the Anti-CaSR antibody levels in plasma samples of HC, R, and LC group. (L) Soluble CaSR levels in plasma samples of HC, R, LC and acute COVID-19 patients. (M) Cumulative data of the plasma ATP in HC, acute, R, and LC groups. Kruskal–Wallis analysis with Dunn’s multiple comparisons test. ns, not significant. Each dot represents a study subject. * p < 0.5, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001.

Journal: Frontiers in Immunology

Article Title: Metabolomic and immune alterations in long COVID patients with chronic fatigue syndrome

doi: 10.3389/fimmu.2024.1341843

Figure Lengend Snippet: Elevated levels of proinflammatory biomarkers in LC patients. (A) Cumulative data comparing the plasma IL-1α, (B) IL-6, (C) TNF-α, (D) IP-10, (E) CRP, (F) SAA, (G) IL-13, and (H) IL-1β, (I) Flt-1, and (J) soluble CD14 measured by ELISA in the plasma of R, LC, and HC groups. (K) Comparing the Anti-CaSR antibody levels in plasma samples of HC, R, and LC group. (L) Soluble CaSR levels in plasma samples of HC, R, LC and acute COVID-19 patients. (M) Cumulative data of the plasma ATP in HC, acute, R, and LC groups. Kruskal–Wallis analysis with Dunn’s multiple comparisons test. ns, not significant. Each dot represents a study subject. * p < 0.5, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001.

Article Snippet: The plasma was subjected to ELISA kit sCD14 (R&D, 383CD-050) ( ) and Anti-CaSR (EAGLE Biosciences).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Pediatric research

Article Title: Ingested soluble CD14 from milk is transferred intact into the blood of newborn rats

doi: 10.1038/pr.2013.225

Figure Lengend Snippet: Biodistribution of intact (>30 kDa, black) and degraded (<30 kDa, white) [ 14 C]sCD14 and a digestible control protein, [ 14 C]BSA, postingestion in 10 d old rats. A, stomach; B, duodenum; C, jejunum; D, ileum; E, large intestine. Rat pups were gavage fed 25 μg of [ 14 C]sCD14 (1.63 ×10 5 dpm/μg) or [ 14 C]BSA (3.44 ×10 4 dpm/μg). Post-euthanasia (0.3, 4 or 8 h), proteins were extracted from organs, size separated using a 30 kDa cut-off spin column and liquid scintillation counted. Data are presented as mean ± SD, and n=3 for each time point. * P <0.05, using a Student’s t -test.

Article Snippet: Human recombinant sCD14 (R&D Systems, Minneapolis, MN) and BSA (Sigma-Aldrich, Oakville, ON, Canada) were labeled with [ 14 C]CH 3 I using 52.9 mCi/mmol (sCD14) or 10 mCi/mmol (BSA) (Perkin Elmer, Waltham, MA and Sigma-Aldrich).

Techniques: Control

Journal: Pediatric research

Article Title: Ingested soluble CD14 from milk is transferred intact into the blood of newborn rats

doi: 10.1038/pr.2013.225

Figure Lengend Snippet: The biodistribution of [ 125 I]sCD14 (A) and a digestible control protein, [ 125 I]BSA (B), post-ingestion in 10 d old rats. Rat pups were gavage fed 25 μg of [ 125 I]sCD14 (12.16 ×10 6 dpm/μg) or [ 125 I]BSA (13.7 ×10 6 dpm/μg). Post-euthanasia (0.3, 4 or 8 h), proteins were extracted from harvested organs, size separated by SDS-PAGE and phosphor-imaged. Stomach (St) and contents (St Con), duodenum (Duo) and contents (Duo Con), jejunum (Je) and contents (Je Con), ileum and contents (Ileum Con), large intestine (L In) and contents (L In Con), liver and blood. Figure is representative of n=3.

Techniques: Control, SDS Page

Journal: Pediatric research

Article Title: Ingested soluble CD14 from milk is transferred intact into the blood of newborn rats

doi: 10.1038/pr.2013.225

Figure Lengend Snippet: Uptake of intact (A, >30 kDa) and degraded (B, <30 kDa) [ 125 I]sCD14 into the blood of 10 d old rat pups. Data are presented as mean ± SD, and n=4. Percent dose fed >30 kDa ( A ) at 0.3 h was significantly less than all subsequent time points (*, P <0.05). Percent dose fed <30 kDa ( B ) at 0.3 h was significantly less than the percent dose fed at 4, 5 and 7 h (*, P <0.05, Student’s t -test).

Techniques:

Journal: Pediatric research

Article Title: Ingested soluble CD14 from milk is transferred intact into the blood of newborn rats

doi: 10.1038/pr.2013.225

Figure Lengend Snippet: Intactness of ingested [ 125 I]sCD14 in the blood of 10 d old rat pups post size separation (< or >30 kDa) visualized by phosphor-imaging (A) and silver staining (B). Intact and non-ingested [ 125 I]sCD14 was used as a positive control. Figure is representative of n=4.

Techniques: Imaging, Silver Staining, Positive Control

Journal: Pediatric research

Article Title: Ingested soluble CD14 from milk is transferred intact into the blood of newborn rats

doi: 10.1038/pr.2013.225

Figure Lengend Snippet: The presence of rat-sCD14 in rat milk, 10 days postpartum, as detected by immunoblotting. Lanes: 1, molecular weight marker; 2, rat milk; 3, human recombinant sCD14 (100 ng, positive control).

Techniques: Western Blot, Molecular Weight, Marker, Recombinant, Positive Control

Journal: Immunology

Article Title: Soluble CD14 is essential for lipopolysaccharide-dependent activation of human intestinal mast cells from macroscopically normal as well as Crohn's disease tissue

doi: 10.1111/imm.12299

Figure Lengend Snippet: Flow cytometric analysis of Toll-like receptor 4 (TLR4) and CD14 surface expression on human intestinal mast cells (hiMC). TLR4 expression (a) was determined by direct immunofluorescence using phycoerythrin-conjugated mouse anti-human CD284 antibody (dark grey profile). Expression of membrane bound CD14 (b) was assessed using FITC-conjugated mouse anti-human CD14 antibody (dark grey profile). Both stainings were compared with unstained hiMC (light grey profile). The data are expressed as cell number versus log fluorescence and are representative of three independent experiments.

Article Snippet: Reagents for performing the experiments were purchased from the following sources: phycoerythrin anti-human CD284 (Toll-like receptor 4; TLR4) from BioLegend (San Diego, CA), FITC anti-human CD14 from BD Biosciences (San Jose, CA); myeloma IgE from Calbiochem by Merck (Darmstadt, Germany); polyclonal anti-human IgE from U. Blank (INSERM, Paris, France); LPS from Escherichia coli serotype O111:B4 from Sigma-Aldrich; recombinant human soluble CD14 (sCD14), recombinant human interferon- γ (IFN- γ ) and recombinant human tumour necrosis factor- α (TNF- α ) from PeproTech; ionomycin and PMA from Sigma-Aldrich; TLR4 signalling inhibitor CLI-095 from InvivoGen (San Diego, CA); RNeasy Mini Kit from Qiagen (Hilden, Germany); SsoFast™ EVAGreen® Supermix from Bio-Rad Laboratories (Munich, Germany); cysteinyl leukotriene (CysLT) EIA kit and IL-1 β EIA kit from Enzo Life Sciences (Lorrach, Germany), CXCL8 Duo Set from R & D Systems (Abingdon, UK), Novex® magnetic Luminex® assay from Invitrogen (Frederick, MD).

Techniques: Expressing, Immunofluorescence, Fluorescence

In the presence of soluble CD14 (sCD14) in ascending concentrations, lipopolysaccharide (LPS) induced the cytokine expression in human intestinal mast cells (hiMC). The hiMC were treated with 1 μg/ml LPS (black bar) or sCD14 (light grey bar) solely, respectively, or with 1 μg/ml LPS plus sCD14 (0·001–1 μg/ml) (dark grey bars) for 3 hr to analyse the mRNA expression of CXCL8 (a), interleukin-1β (IL-1β) (b), IL-6 (c), CCL2 (d), CCL3 (e), and CCL4 (f). As positive control for determination of hiMC ability to induce cytokine mRNA expression, cells were treated with myeloma IgE for 90 min and afterwards stimulated with anti-human IgE for 90 min for cross-linking of high-affinity IgE receptor (FcεRI CL, black striped bar). Data represent mean ± SEM of five or six independent experiments. *P < 0·05 and **P < 0·001 indicate significant differences from hiMC treated with LPS (1 μg/ml) alone (black bar).

Journal: Immunology

Article Title: Soluble CD14 is essential for lipopolysaccharide-dependent activation of human intestinal mast cells from macroscopically normal as well as Crohn's disease tissue

doi: 10.1111/imm.12299

Figure Lengend Snippet: In the presence of soluble CD14 (sCD14) in ascending concentrations, lipopolysaccharide (LPS) induced the cytokine expression in human intestinal mast cells (hiMC). The hiMC were treated with 1 μg/ml LPS (black bar) or sCD14 (light grey bar) solely, respectively, or with 1 μg/ml LPS plus sCD14 (0·001–1 μg/ml) (dark grey bars) for 3 hr to analyse the mRNA expression of CXCL8 (a), interleukin-1β (IL-1β) (b), IL-6 (c), CCL2 (d), CCL3 (e), and CCL4 (f). As positive control for determination of hiMC ability to induce cytokine mRNA expression, cells were treated with myeloma IgE for 90 min and afterwards stimulated with anti-human IgE for 90 min for cross-linking of high-affinity IgE receptor (FcεRI CL, black striped bar). Data represent mean ± SEM of five or six independent experiments. *P < 0·05 and **P < 0·001 indicate significant differences from hiMC treated with LPS (1 μg/ml) alone (black bar).

Techniques: Expressing, Positive Control

Time-dependent expression and release of CXCL8 (a, b) and interleukin (IL)-1β (c) by human intestinal mast cells (hiMC) upon administration of lipopolysaccharide (LPS) in combination with soluble CD14 (sCD14). The hiMC were incubated with 1 μg/ml LPS (black bar) or sCD14 (light grey bar) solely for 6 hr, or with LPS plus sCD14 (each 1 μg/ml) (dark grey bars) for 3 hr, 6 hr and 12 hr to analyse protein production of CXCL8 (a) and IL-1β (c) in cell lysate and release of CXCL8 (b) in supernatants. In case of CXCL8 protein production, IgE receptors were cross-linked with myeloma IgE (90 min) and anti-human IgE for 6 hr (FcεRI CL, black striped bar) as stimulation control. For CXCL8 in cell lysates (a) and supernatants (b), data represent mean ± SEM of three independent experiments by treatments with LPS plus sCD14 (dark grey bars) and cross-linking of high-affinity IgE receptor (black striped bar). No CXCL8 protein was measured in cell lysates and supernatants of hiMC without any stimulation (a, b) and treated exclusively with LPS (b). In the case of cytosolic CXCL8 protein content (a), data represent mean ± SEM of two independent experiments for sole incubation with LPS (black bar) and sCD14 (light grey bar), respectively. After treatment of hiMC with sCD14 only (light grey bar), a single measurement was quantified and is represented for CXCL8 concentration in cell supernatants (b). According to the missing values concerning cytosolic content and release of CXCL8 by LPS treatment alone, no statistical analysis with the paired t-test was performed for these data. In case of cytosolic release of IL-1β and its precursor (c), data represent mean ± SEM of four independent experiments. *P < 0·05 indicate significant differences to hiMC treated with LPS [1 μg/ml] solely (black bar).

Journal: Immunology

Article Title: Soluble CD14 is essential for lipopolysaccharide-dependent activation of human intestinal mast cells from macroscopically normal as well as Crohn's disease tissue

doi: 10.1111/imm.12299

Figure Lengend Snippet: Time-dependent expression and release of CXCL8 (a, b) and interleukin (IL)-1β (c) by human intestinal mast cells (hiMC) upon administration of lipopolysaccharide (LPS) in combination with soluble CD14 (sCD14). The hiMC were incubated with 1 μg/ml LPS (black bar) or sCD14 (light grey bar) solely for 6 hr, or with LPS plus sCD14 (each 1 μg/ml) (dark grey bars) for 3 hr, 6 hr and 12 hr to analyse protein production of CXCL8 (a) and IL-1β (c) in cell lysate and release of CXCL8 (b) in supernatants. In case of CXCL8 protein production, IgE receptors were cross-linked with myeloma IgE (90 min) and anti-human IgE for 6 hr (FcεRI CL, black striped bar) as stimulation control. For CXCL8 in cell lysates (a) and supernatants (b), data represent mean ± SEM of three independent experiments by treatments with LPS plus sCD14 (dark grey bars) and cross-linking of high-affinity IgE receptor (black striped bar). No CXCL8 protein was measured in cell lysates and supernatants of hiMC without any stimulation (a, b) and treated exclusively with LPS (b). In the case of cytosolic CXCL8 protein content (a), data represent mean ± SEM of two independent experiments for sole incubation with LPS (black bar) and sCD14 (light grey bar), respectively. After treatment of hiMC with sCD14 only (light grey bar), a single measurement was quantified and is represented for CXCL8 concentration in cell supernatants (b). According to the missing values concerning cytosolic content and release of CXCL8 by LPS treatment alone, no statistical analysis with the paired t-test was performed for these data. In case of cytosolic release of IL-1β and its precursor (c), data represent mean ± SEM of four independent experiments. *P < 0·05 indicate significant differences to hiMC treated with LPS [1 μg/ml] solely (black bar).

Techniques: Expressing, Incubation, Concentration Assay

Down-regulation of induced mRNA expression of CXCL8 (a) and interleukin (IL)-1β (b) due to administration of Toll-like receptor 4 (TLR4) inhibitor before treatment with lipopolysaccharide (LPS) and soluble CD14 (sCD14). In advance of LPS/sCD14-stimulation, human intestinal mast cells (hiMC) were treated with the TLR4 inhibitor CLI-095 at concentrations of 0·1 and 1 μm for 60 min and afterwards incubated with LPS combined with sCD14 (each 1 μg/ml) (dark grey bars) for 3 hr to determine mRNA expression. Data represent mean ± SEM of six independent experiments. *P < 0·05 indicate significant differences between groups.

Journal: Immunology

Article Title: Soluble CD14 is essential for lipopolysaccharide-dependent activation of human intestinal mast cells from macroscopically normal as well as Crohn's disease tissue

doi: 10.1111/imm.12299

Figure Lengend Snippet: Down-regulation of induced mRNA expression of CXCL8 (a) and interleukin (IL)-1β (b) due to administration of Toll-like receptor 4 (TLR4) inhibitor before treatment with lipopolysaccharide (LPS) and soluble CD14 (sCD14). In advance of LPS/sCD14-stimulation, human intestinal mast cells (hiMC) were treated with the TLR4 inhibitor CLI-095 at concentrations of 0·1 and 1 μm for 60 min and afterwards incubated with LPS combined with sCD14 (each 1 μg/ml) (dark grey bars) for 3 hr to determine mRNA expression. Data represent mean ± SEM of six independent experiments. *P < 0·05 indicate significant differences between groups.

Techniques: Expressing, Incubation

Journal: Immunology

Article Title: Soluble CD14 is essential for lipopolysaccharide-dependent activation of human intestinal mast cells from macroscopically normal as well as Crohn's disease tissue

doi: 10.1111/imm.12299

Figure Lengend Snippet: In the presence of soluble CD14 (sCD14) in ascending concentrations, lipopolysaccharide (LPS) induced the cytokine expression in human intestinal mast cells (hiMC) from patients with Crohn's disease. The hiMC were treated with 1 μg/ml LPS (black bar) or sCD14 (light grey bar) solely, respectively, or with 1 μg/ml LPS plus sCD14 (0·001–1 μg/ml) (dark grey bars) for 3 hr to analyse the mRNA expression of CXCL8 (a), interleukin-1β (IL-1β) (b), IL-6 (c), CCL2 (d), CCL3 (e), and CCL4 (f). As positive control for determination of hiMC ability to induce cytokine mRNA expression, cells were treated with myeloma IgE for 90 min and afterwards stimulated with anti-human IgE for 90 min for cross-linking of high-affinity IgE receptor (FcεRI CL, black striped bar). Data represent mean ± SEM of four to six independent experiments. *P < 0·05 indicates significant differences from hiMC treated with LPS (1 μg/ml) solely (black bar).

Techniques: Expressing, Positive Control

Comparison of CXCL8 mRNA (a) and protein (b) expression in human intestinal mast cells (hiMC) isolated from macroscopically normal tissue specimens (Co-MC) and from patients with Crohn's disease (CD-MC). For analysing mRNA expression of CXCL8 (a, b), Co-MC and CD-MC were treated with 1 μg/ml lipopolysaccharide (LPS; black bar) or soluble CD14 (sCD14) (light grey bar) solely for 6 hr, or with LPS plus sCD14 (each 1 μg/ml) (dark grey bars) for 90 min, 3 hr and 6 hr. As positive control for determination of the ability of hiMC to induce cytokine mRNA expression, cells were treated with myeloma IgE for 90 min and afterwards stimulated with anti-human IgE for 90 min for cross-linking of high-affinity IgE receptor (FcεRI CL, black striped bar). Data represent mean ± SEM of four or five independent experiments. *P < 0·05 indicates significant differences from hiMC treated with LPS (1 μg/ml) solely (black bar). Cytosolic content (c) and release of CXCL8 (d) were determined in cell lysate and supernatants, respectively. For this purpose, Co-MC (white bars) and CD-MC (black bars) were treated with LPS in combination with sCD14 (each 1 μg/ml) for 6 hr or with myeloma IgE for 90 min and afterwards stimulated with anti-human IgE for 6 hr for cross-linking of high-affinity IgE receptor (FcεRI CL), respectively. Data represent mean ± SEM of four to six independent experiments; a: P < 0·05 compared with Control (6 hr) of Co-MC, b: P < 0·05 compared with Control (6 hr) of CD-MC.

Journal: Immunology

Article Title: Soluble CD14 is essential for lipopolysaccharide-dependent activation of human intestinal mast cells from macroscopically normal as well as Crohn's disease tissue

doi: 10.1111/imm.12299

Figure Lengend Snippet: Comparison of CXCL8 mRNA (a) and protein (b) expression in human intestinal mast cells (hiMC) isolated from macroscopically normal tissue specimens (Co-MC) and from patients with Crohn's disease (CD-MC). For analysing mRNA expression of CXCL8 (a, b), Co-MC and CD-MC were treated with 1 μg/ml lipopolysaccharide (LPS; black bar) or soluble CD14 (sCD14) (light grey bar) solely for 6 hr, or with LPS plus sCD14 (each 1 μg/ml) (dark grey bars) for 90 min, 3 hr and 6 hr. As positive control for determination of the ability of hiMC to induce cytokine mRNA expression, cells were treated with myeloma IgE for 90 min and afterwards stimulated with anti-human IgE for 90 min for cross-linking of high-affinity IgE receptor (FcεRI CL, black striped bar). Data represent mean ± SEM of four or five independent experiments. *P < 0·05 indicates significant differences from hiMC treated with LPS (1 μg/ml) solely (black bar). Cytosolic content (c) and release of CXCL8 (d) were determined in cell lysate and supernatants, respectively. For this purpose, Co-MC (white bars) and CD-MC (black bars) were treated with LPS in combination with sCD14 (each 1 μg/ml) for 6 hr or with myeloma IgE for 90 min and afterwards stimulated with anti-human IgE for 6 hr for cross-linking of high-affinity IgE receptor (FcεRI CL), respectively. Data represent mean ± SEM of four to six independent experiments; a: P < 0·05 compared with Control (6 hr) of Co-MC, b: P < 0·05 compared with Control (6 hr) of CD-MC.

Techniques: Expressing, Isolation, Positive Control

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human recombinant scd14 (R&D Systems)

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